β-Adrenergic receptor signaling, desensitization, and downregualtion are fundamental mechanisms that contribute to both normal and altered myocar dial function. The development of pharmacological and biochemical assays has provided the ability to measure alterations b ...

The sarcoplasmic reticulum Casn+2 ATPase (SERCA) plays a pivotal role in calcium cycling and the beat-to-beat function of the heart. Recent studies have shown that decreased expression and activity of SERCA are associated with end-stage heart failure in humans and in experimental animal m ...

This chapter describes the methods required for overexpression of the angiotensin II type I receptor (AT1) in cardiomyocytes of transgenic rats. This includes cloning of the transgenic construct consisting of the α-myosin heavy chain (MHC) promoter, the human AT1 cDNA and SV40 T-antigen s ...

Diabetic cardiomyopathy is a common chronic complication leading to height ened risk of heart failure among diabetic patients. In this chapter, we describe the methods for maintenance and breeding of two diabetic animal models, OVE26 and Agouti mice, for type 1 and type 2 diabetes, respectiv ...

Endothelial cells function as a highly regulated barrier between blood and interstitium. They play a central role in the regulation of vascular permeability by controlling the passage of liquid and nutrients as well as the transit of white blood cells (1,2). The endothelium is involved in the in ...

Airway smooth muscle (ASM) cells may be considered an important modulator of airway inflammatory responses, since they synthesize cytokines, chemokines, growth factors and metalloproteinases as well as support the adhesion of activated T lymphocytes. The study of the cellular and m ...

Mast cells were once thought to represent a single population of highly granulated secretory cells. However, with the development of mast cell isolation techniques for a diverse range of tissues, it became apparent that mast cells from different species and those from different tissues wi ...

Within the human lung, macrophages can be found in the pleura, interstitium, alveoli, airways, vasculature, and walls of the bronchi and bronchiols. This distribution does not simply reflect the ubiquitous nature of these cells, as the macrophages found at these different sites show subtle d ...

Neutrophils are manufactured in the bone marrow from stem cell precursors (myeloblasts) by a series of cell divisions. In healthy individuals only the mature neutrophils enter peripheral blood. They are the most numerous leukocyte subgroup, constituting half of the circulating white ...

There is increasing evidence that T lymphocytes play a central role in regulating both the initial and chronic inflammatory cascades of allergic asthma (1,2) and can also regulate baseline airway responsiveness in mouse models of asthma (3). In particular, activation of antigen-specif ...

Granulocytes play an important role in the host defense against invading microorganisms such as bacteria and fungi (1,2). For the killing reaction, these cells have an extensive machinery of cytotoxic effector mechanisms including phagocytosis, production of toxic oxygen metabo ...

Numerous techniques have been developed to measure gene expression in tissues and cells. These include coupled reverse transcription and polymerase chain reaction amplification (RT-PCR), Northern blot (see Chapter 20), in situ hybridization (see Chapter 21), RNase protection as ...

Messenger (m)RNAs represent the information-carrying intermediates for protein synthesis. Northern blotting gives a snapshot of the relative abundance of an RNA transcript at a set time point by measuring the steady-state levels of an RNA transcript. The steady-state level of a partic ...

In situ hybridization (ISH) (also called “hybridization histochemistry” or “hybridization cytology”) was first described in 1969 by Gall and Pardue who used the technique to localize ribosomal DNA in Xenopus oocytes (1). In contrast to other techniques of hybridization (i.e., Northern ...

Hypersecretion of airway mucus is characteristic of several severe lung diseases, particularly those involving chronic inflammation such as asthma, chronic obstructive pulmonary disease (COPD), and cystic fibrosis (CF) (1). Mucins are the major macromolecular component of mu ...

In chronic inflammatory diseases, the expression of multiple genes, including those for cytokines, chemokines, adhesion molecules, receptors, and inflammatory enzymes, is often upregulated. The problem for many academic or industrial scientists is to elucidate the mechanisms ...

Flow cytometry is an invaluable tool for the analysis of leukocyte populations in inflammation. Flow cytometers, particularly those designed purely for analysis rather than cell sorting, have now become user-friendly machines and are commonplace in many laboratories. Any cell type ...

Airway inflammation is currently the subject of intense research interest concerning both the nature of the inflammatory cells, proteins, and cytokines present. These data are being used to define and assess the severity, cause, prognosis, and response to treatment of airway inflammat ...

During the Phase I/II assessment of new therapies with the potential to suppress eosinophil and neutrophil inflammation, there is a need to assess the peripheral blood pharmacokinetic (PK) and pharmacodynamic (PD) profiles of the drug. This has relevance in respiratory disease since dr ...

A large component of airway inflammatory disease is the recruitment of activated leukocytes (primarily eosinophils and T lymphocytes) from the lung vasculature into the bronchial walls resulting in lung edema. Ultimately, many of the infiltrating leukocytes progress across the a ...